@article {Roffay2022.09.28.509885, author = {Chlo{\'e} Roffay and Juan Manuel Garc{\'\i}a-Arcos and Pierrik Chapuis and Javier L{\'o}pez-Andarias and Falk Schneider and Adai Colom and Caterina Tomba and Ilaria Di Meglio and Valentin Dunsig and Stefan Matile and Aur{\'e}lien Roux and Vincent Mercier}, title = {Technical insights into fluorescence lifetime microscopy of mechanosensitive Flipper probes}, elocation-id = {2022.09.28.509885}, year = {2023}, doi = {10.1101/2022.09.28.509885}, publisher = {Cold Spring Harbor Laboratory}, abstract = {Measuring forces within living cells remains a technical challenge. We developed hydrophobic mechanosensing fluorescent probes called Flippers, whose fluorescence lifetime depends on lipid packing and can report on membrane tension. Here, we describe technical optimization of the probe imaging, and diverse characterizations in various biological and in vitro systems. We provide a guideline to measure biophysical parameters of cellular membranes by FLIM microscopy with Flipper probes, providing evidences that flippers can report long range forces in cells, tissues and organsi.Competing Interest StatementThe authors have declared no competing interest.}, URL = {https://www.biorxiv.org/content/early/2023/01/31/2022.09.28.509885}, eprint = {https://www.biorxiv.org/content/early/2023/01/31/2022.09.28.509885.full.pdf}, journal = {bioRxiv} }